Comparative Analysis of Some DNA Extraction Kits used for Molecular Analysis of Iron Ore Samples

Ekpa E*, Asmau M, Onwurah I.N.E, Ezeanyika L.U.S, Etuk-Udo,G

Bioleaching simply refers to the microbial-catalyzed process of conversion of insoluble metals into their soluble forms. As an applied biotechnology, it represents an extremely interesting field of research where genomic and proteomic techniques can be used in terms of knowledge development, but more in terms of process design, control, and optimization. Good quality DNA is a prerequisite for all experiments requiring DNA manipulation. DNA isolation is often the most fundamental and crucial in the entire process. As a result of different DNA isolation kits constantly becoming available, it has become imperative to compare their efficacies. This study compared some popular isolation kits used on bioleaching microorganisms from an Iranian Iron mine with that of Zymo research kit used on Agbaja Iron stones of Nigeria. The Iron mine of Agbaja in kogi State possess similar characteristics with those of the Iranian Iron mine-hence the choice of the comparative protocols. Modified CTAB method (Sigma) and kits of the QIAmp Investigator (Qiagen) and Accuprep Genomic DNA (Bioneer) were used to compare to that of ZR Genomic DNA (Zymo Research) used in this work. Spectrophotometric results showed that the highest concentration of DNA from 100µl of bioleaching bacteria culture was obtained by Qiagen kit (165.50μg/μl) followed by Zymo kit (50.40μg/μl), Bioneer kit (34.33μg/μl) and CTAB (23.65μg/μl). The Bioneer test took a total of 40 minutes to isolate bacterial DNA, followed by Zymo kit, 45 minutes, Qiagen, 50 minutes, and the CTAB 60minutes. These preliminary findings may serve as future baseline information in helping laboratories/Researchers make appropriate decisions in process optimization for isolating quality DNA especially from bioleaching microorganisms.

A Volumetric Assessment with a 3D Scanner of an Eye Reconstruction Fabricated Using the Traditional Handmade Method. A Comparative Pilot Case Study

Sonia Regina Bordin-Aykroyd, Brito Dias R, Lynch E , Coto N P

Purpose: Precision in anatomical details and the quantification of volume loss is important in bucco-maxillofacial prosthetic reconstruction for the accurate symmetrical rehabilitation of patients with bucco maxillo-facial loss or malformations. The objective of this case study was to determine if a prosthetic eye made by the traditional handmade system, using hydrocolloid impression, had the same size as the natural eye and evaluate the role of 3D scanners in this process. In order to determine if the two eyes had the same size, a 3D scanner was used to measure and compare the volume of the depressions created by theses eyes after hydrocolloid impressions where taken. These depression measurements enabled us to calculate the thickness of the eyes, quantifying the accuracy of the traditional confection method.

Materials &Methods: The comparison was made, after randomly assigning one patient out of 100, who had ocular prosthesis previously constructed in the traditional way, with the use of hydrocolloid material for the impression of the anophthalmic cavity. Stone models were made from hydrocolloid molds of the surface of the eyelids containing: the natural eye, the prosthetic eye, the eyelid of the empty anophthalmic cavity and the mold of the prosthetic eye alone. Following this, measuring tests using a 3D scanner were made and the results were compared.

Results: The surface volume of the empty anophthalmic cavity eyelid was 23% smaller than that of the natural eye; while, with the prosthetic eye in place, it was 8% smaller. This indicated that in order to have a closer appearance to the natural eye and a more natural contour, the prosthetic eye would need to be increased in volume. The volume measurement of the prosthetic eye was made using the 3D scanner and was 2750 mm3. A further measurement, using the computer model, suggested that for the eyelids surface volume to be the same, the prosthetic eye would need to be increased by 2,5 to 5 %, in thickness, depending where the increment was made.

Conclusion: These findings suggest that the traditional handmade method of constructing prosthetic eyes, with impressions from hydrocolloid materials, can produce a series of discrepancies during the confection process, and can lead to a significant difference in size from that of the natural eye. 3D scanners could offer a valuable impression method for bucco maxillo-facial reconstruction, as it allows for the delineation of the area of interest to be rebuilt.

Volume 2 Issue 2

A Handy MCR Modification for PCR Cloning into Plasmid Vectors

Anastasia Sadova, Kirill Netchvolodov, Natalia Kupriyanova*

There exist a lot of vectors for a wide range of experiments, and new modified plasmids appear every year. Most of them contain different Multiple Cloning Regions (MCR)s depending on specificity of the experiments. The main problem of such the trial-and-error method is that a lot of MCRs contain a limited number of Restriction Sites (RS), which often makes the process of subcloning arduous. Here, we elaborated design of the MCR for a GeneClip vector that can be used both for cloning PCR products and transferring the insert from one plasmid to the others. So, everybody can now construct MCRs for their own vectors using this manual.